This invention is directed to compounds and their preparation, to pharmaceutical compositions containing the compounds and to their use in the treatment of physiological conditions capable of being modulated by agonist or antagonist activity on parathyroid hormone receptors. More particularly, this invention is directed to peptide parathyroid hormone analogs and peptide parathyroid hormone related protein analogs.
Human parathyroid hormone (hPTH) is an 84 amino acid protein which is a major regulator of calcium homeostasis. Parathyroid hormone-related protein (hPTHrP) is a 139 to 171 amino acid protein with N-terminal homology to hPTH. The N-terminal fragments of hPTH and hPTHrP, particularly those consisting of amino acids 1-34, retain the full biological activity of the parent hormone.
hPTH(1-34) has the following amino acid sequence:
Ser-Val-Ser-Glu-Ile-Gln-Leu-Met-His-Asn-Leu-Gly-Lys-His-Leu-Asn-Ser-Met-
Glu-Arg-Val-Glu-Trp-Leu-Arg-Lys-Lys-Leu-Gln-Asp-Val-His-Asn-Phe. (SEQ ID NO: 1)
hPTHrP has the following amino acid sequence:
Ala-Val-Ser-Glu-His-Gln-Leu-Leu-His-Aso-Lys-Gly-Lys-Ser-Ile-(Gln-Aso-Leu-
Arg-Arg-Arg-Phe-Phe-Leu-His-His-Leu-Ile-Ala-Glu-Ile-His-Thr-Ala. (SEQ ID NO: 2)
The biological activity of hPTH is reflected in the activation of two secondary messenger systems: G-protein coupled adenylyl cyclase (AC) and G-protein coupled and uncoupled protein kinase C (PKC) activity. The N-terminal fragments hPTH(1-34)OH and hPTH(1-31)NH2 have been demonstrated to be anabolic with respect to bone formation in humans and ovariectomized rats, respectively. This increase in bone growth has been demonstrated to be coupled with stimulation of adenylyl cyclase activity. Analogs of these N-terminal fragments have significant therapeutic potential for the treatment of physiological conditions associated with bone cell calcium regulation including hypocalcemia; osteoporosis; osteopenia; and disorders associated with osteoporosis and osteopenia such as hyperparathyroidism, hypoparathyroidism, and Cushings syndrome; glucocorticoid- and immunosuppressant-induced osteopaenia; and bone fracture and bone refracture repair.
It has also been established that deletion of up to six amino acid residues from the N-terminus of hPTH(1-34) markedly decreases the resulting analog""s ability to stimulate adenylyl cyclase while having little effect on receptor binding. Thus, analogs of hPTH(1-34) truncated by up to six amino acid residues at the N-teminus inhibit the action of PTH and are useful in the treatment of disorders characterized by an excess of PTH such as hyperparathyrodism and hyperparathyrodism-related hypercalcemia crisis, hypercalcemia of malignancy, renal failure and hypertension.
Acyclic analogs of hPTH(1-27) to (1-34) are disclosed in U.S. Pat. No. 4,086,196. Acyclic analogs of hPTH(1-34) and hPTHrP (1-34) are disclosed in U.S. Pat. No. 5,589,452. [Nle8, Nle18, Tyr34, or Phe34]hPTH(1-34) are disclosed in U.S. Pat. No. 4,656,250. [Nle8, Nle18, Tyr34]hPTH(1-34) and N-truncated derivatives thereof are disclosed in U.S. Pat. Nos. 4,771,124 and 4,423,037. Other acyclic analogs of PTH(1-34) are disclosed in U.S. Pat. Nos. 5,723,577 and 5,434,246, WO 97/02834, EPA 561 412-A1, EPA 748,817-A2, WO-94/02510, WO9603437, and WO951 1988-A 1. Analogs of hPTH(1-28)NH2 to hPTH(1-31)NH2 and [Leu27]hPTH(1-28)NH2 to [Leu27]hPTH(1-33)NH2 are decribed in U.S. Pat. No. 5,556,940. Acyclic antagonists of the PTH receptor including N-terminally-truncated analogs of PTH are disclosed in U.S. Pat. Nos. 5,446,130, 5,229,489, 4,771,124 and 4,423,037.
Cyclic and bicyclic analogs of hPTH and hPTHrP have been disclosed. Cyclo(Lys26-Asp30)[Leu27]hPTH(1-34)NH2 and cyclo(Lys27-Asp30)hPTH(1-34)NH2 are disclosed in U.S. Pat. No. 5,556,940. Cyclo(Lys26-Asp30)[Leu27]hPTH(1-31)NH2, cyclo(Glu22-Lys26)[Leu27]hPTH(1-31)NH2, and cyclo(Lys27-Asp30)hPTH(1-31)NH2 are decribed by Barbier, et al., J. Med. Chem. 1997, 40, 1373. Monocyclic and bicyclic derivatives of hPTH(1-34) or hPTHrP(1-34) are disclosed in patent documents WO 96/40193. DE19508672-A1, and by A. Bisello. et al., in Biochemistry 1997, 36, 3293. Cyclo(Lys13-Asp17)hPTHrP(7-34)NH2, a potent antagonist of the PTH rcceptor, is disclosed by M. Chorev, et al., Biochemistry 1991, 30, 5698. Also, Kanmera, et al., has described a series of amide-containing analogs of hPTHrP, Peptide Chemistry 1993: Okada, Y., ed.; Protein Research Foundation, Osaka, 1994, 321-324.xe2x80x9d
This invention is directed to a cyclic peptide compound of formula I
Xxe2x80x94A10xe2x80x94A11xe2x80x94A12xe2x80x94A13xe2x80x94A14xe2x80x94A15xe2x80x94A16xe2x80x94A17xe2x80x94A18xe2x80x94A19xe2x80x94A20xe2x80x94A21xe2x80x94A22xe2x80x94A23xe2x80x94A24xe2x80x94A25xe2x80x94A26xe2x80x94A27xe2x80x94Yxe2x80x83xe2x80x83I
or a pharmaceutically acceptable salt or prodrug thereof wherein
X is selected from the group consisting of
(a) R1axe2x80x94A0xe2x80x94A1xe2x80x94A2xe2x80x94A3xe2x80x94A4xe2x80x94A5xe2x80x94A6xe2x80x94A7xe2x80x94A8xe2x80x94A9xe2x80x94,
(b) R1axe2x80x94A2xe2x80x94A3xe2x80x94A4xe2x80x94A5xe2x80x94A6xe2x80x94A7xe2x80x94A8xe2x80x94A9xe2x80x94,
(c) R1bxe2x80x94A3xe2x80x94A4xe2x80x94A5xe2x80x94A6xe2x80x94A7xe2x80x94A8xe2x80x94A9xe2x80x94,
(d) R1axe2x80x94A4xe2x80x94A5xe2x80x94A6xe2x80x94A7xe2x80x94A8xe2x80x94A9xe2x80x94,
(e) R1axe2x80x94A5xe2x80x94A6xe2x80x94A7xe2x80x94A8xe2x80x94A9xe2x80x94,
(f) R1axe2x80x94A6xe2x80x94A7xe2x80x94A8xe2x80x94A9xe2x80x94,
(g) R1axe2x80x94A7xe2x80x94A8xe2x80x94A9xe2x80x94,
(h) R1axe2x80x94A8xe2x80x94A9xe2x80x94,
(i) R1axe2x80x94A9xe2x80x94, and
(j) R1axe2x80x94;
Y is selected from the group consisting of
(a) xe2x80x94R3,
(b) xe2x80x94A28xe2x80x94R3,
(c) xe2x80x94A28xe2x80x94A29xe2x80x94R3,
(d) xe2x80x94A28xe2x80x94A29xe2x80x94A30xe2x80x94R3,
(e) xe2x80x94A28xe2x80x94A29xe2x80x94A30xe2x80x94A31xe2x80x94R3,
(f) xe2x80x94A28xe2x80x94A29xe2x80x94A30xe2x80x94A31xe2x80x94A32xe2x80x94R3,
(g) xe2x80x94A28xe2x80x94A29xe2x80x94A30xe2x80x94A31xe2x80x94A32xe2x80x94A33xe2x80x94R3, and
(h) xe2x80x94A28xe2x80x94A29xe2x80x94A30xe2x80x94A31xe2x80x94A32xe2x80x94A33xe2x80x94A34xe2x80x94R3;
R1a is H, alkyl, aralkyl or xe2x80x94COR2;
R1b is R1a or a group of formula 
R2 is alkyl, alkenyl, alkynyl, aryl or aralkyl;
R3 is a group of formula A35xe2x80x94OR4 or A35xe2x80x94NR4R5;
R4 and R5 are independently H or lower alkyl;
R6 and R9 are independently H or alkyl;
R7 is alkyl;
R8 is H, alkyl or COR2;
R10 is H or halogen;
R11 is alkyl or aralkyl;
m is 1, 2 or 3;
n is 3 or 4;
A0 is absent or a peptide of from one to six amino acid residues;
A1 is Ser, Ala, Gly or D-Pro, or an equivalent amino acid thereof;
A2 is Ala, Val or Gly, or an equivalent amino acid thereof;
A3 is Ala, Ser, Gly or D-Pro, or an equivalent amino acid thereof;
A4 is Glu, Ala or Gly, or an equivalent amino acid thereof;
A5 is Ile, His, Ala or Gly, or an equivalent amino acid thereof;
A6 is Ala, Gln, Gly or D-Pro, or an equivalent amino acid thereof;
A7 is Ala, Leu, Gly, or an equivalent amino acid thereof;
A8 is Leu, Nle, Gly or D-Pro, or an equivalent amino acid thereof;
A9 is His, Ala, D-Pro or Gly, or an equivalent amino acid thereof;
A10 is Ala, Asn, Asp, Cys, homo-Cys, Glu, Gly, Lys, Orn, Ser, Thr, D-Pro, xe2x80x94NHCH(CH2)mNH2)COxe2x80x94 or xe2x80x94NHCH[(CH2)nCO2H]COxe2x80x94;
A11 is Ala, Gly, Leu or Lys, or an equivalent amino acid thereof;
A12 is Ala or Gly, or an equivalent amino acid thereof;
A13 is Ala, Asn, Asp, Cys, homo-Cys, Glu, Gly, Lys, Orn, Ser, Thr, xe2x80x94NHCH(CH2)mNH2)COxe2x80x94 or xe2x80x94NHCH[(CH2)nCO2H]COxe2x80x94;
A14 is Ala, Asn, Asp, Cys, homo-Cys, Glu, Gly, His, Lys, Orn, Ser, Thr, D-Pro, xe2x80x94NHCH(CH2)mNH2)COxe2x80x94 or xe2x80x94NHCH[(CH2)nCO2H]COxe2x80x94;
A15 is Ala, Gly, Ile, D-Pro or Leu, or an equivalent amino acid thereof;
A16 is Asn, Ala, Gly, D-Pro or Gln, or an equivalent amino acid thereof;
A17 is Ala, Asn, Asp, Cys, homo-Cys, Glu, Gly, Lys, Orn, Ser, Thr, D-Pro, xe2x80x94NHCH(CH2)mNH2)COxe2x80x94 or xe2x80x94NHCH[(CH2)nCO2H]COxe2x80x94;
A18 is Asp, Cys, homo-Cys, Glu, His, Leu, Lys, Or, Nle, Ser, Thr, xe2x80x94NHCH(CH2)mNH2)COxe2x80x94 or xe2x80x94NHCH[(CH2)nCO2H]COxe2x80x94;
A19 is Arg or Glu, or an equivalent amino acid thereof;
A20 is Arg or an equivalent amino acid thereof;
A21 is Arg, Asp, Cys, homo-Cys, Glu, Lys, Orn, Ser, Thr, Val, xe2x80x94NHCH(CH2)mNH2)COxe2x80x94 or xe2x80x94NHCH[(CH2)nCO2H]COxe2x80x94;
A22 is Asp, Cys, homo-Cys, Glu, His, Lys, Orn, Phe, Ser, Thr, xe2x80x94NHCH(CH2)mNH2)COxe2x80x94 or xe2x80x94NHCH[(CH2)nCO2H]COxe2x80x94;
A23 is Leu, Phe or Trp, or an equivalent amino acid thereof;
A24 is Leu or an equivalent amino acid thereof;
A25 is Arg, Asp, Cys, homo-Cys, Glu, His, Lys, Orn, D-Pro, Ser, Thr, xe2x80x94NHCH(CH2)mNH2)COxe2x80x94 or xe2x80x94NHCH[(CH2)nCO2H]COxe2x80x94;
A26 is Asp, Cys, homo-Cys, Glu, His, Lys, Orn, Ser, Thr, xe2x80x94NHCH(CH2)mNH2)COxe2x80x94 or xe2x80x94NHCH[(CH2)nCO2H]COxe2x80x94;
A27 is Leu or Lys, or an equivalent amino acid thereof;
A28 is Ile or Leu, or an equivalent amino acid thereof;
A29 is Ala, Asp, Cys, homo-Cys, Glu, Gln, Lys, Orn, Ser, Thr, xe2x80x94NHCH(CH2)mNH2)COxe2x80x94 or xe2x80x94NHCH[(CH2)nCO2H]COxe2x80x94;
A30 is Asp, Cys, homo-Cys, Glu, Gly, Lys, Orn, Ser, Thr, xe2x80x94NHCH(CH2)mNH2)COxe2x80x94 or xe2x80x94NHCH[(CH2)nCO2H]COxe2x80x94;
A31 is Ile, Leu or Val, or an equivalent amino acid thereof;
A32 is His, or an equivalent amino acid thereof;
A33 is Asn or Thr, or an equivalent amino acid thereof;
A34 is Ala or Phe, or an equivalent amino acid thereof;
A35 is absent or a peptide of from 1 to 4 amino acids; and
the side chains of at least one of the following pairs of amino acid residues, A10 and A14, A13 and A17, A14 and A18, A17 and A21, A18 and A22, A21 and A25, A25 and A29 and A26 and A30 are linked through an amide, ester, disulfide or lanthionine bond to form a bridge, and the side chain of each of the following amino acid residues, A10, A13, A14, A17, A18, A21, A22, A25, A26, A29, and A30 contributes, at most, to the formation of a single bridge; provided that when the side chains of the following pairs of amino acid acid residues, A13 and A17 or A26 and A30 are linked through an amide, disulfide or lanthionine bond to form a bridge, then the side chains of at least one of the following pairs of amino acid residues, A10 and A14, A14 and A18, A17 and A21, A18 and A22, A21 and A25 and A25 and A29 are also linked through an amide, ester, disulfide or lanthionine bond.
In another aspect, this invention is directed to a peptide compound of formula II
Xxe2x80x94A10xe2x80x94A11xe2x80x94A12xe2x80x94A13xe2x80x94A14xe2x80x94A15xe2x80x94A16xe2x80x94A17xe2x80x94A18xe2x80x94A19xe2x80x94A20xe2x80x94A21xe2x80x94A22xe2x80x94A23xe2x80x94A24xe2x80x94A25xe2x80x94A26xe2x80x94A27xe2x80x94Yxe2x80x83xe2x80x83II
or a pharmaceutically acceptable salt or prodrug thereof wherein
X is selected from the group consisting of
(a) R1axe2x80x94A0xe2x80x94A1xe2x80x94A2xe2x80x94A3xe2x80x94A4xe2x80x94A5xe2x80x94A6xe2x80x94A7xe2x80x94A8xe2x80x94A9xe2x80x94,
(b) R1axe2x80x94A2xe2x80x94A3xe2x80x94A4xe2x80x94A5xe2x80x94A6xe2x80x94A7xe2x80x94A8xe2x80x94A9xe2x80x94,
(c) R1bxe2x80x94A3xe2x80x94A4xe2x80x94A5xe2x80x94A6xe2x80x94A7xe2x80x94A8xe2x80x94A9xe2x80x94,
(d) R1axe2x80x94A4xe2x80x94A5xe2x80x94A6xe2x80x94A7xe2x80x94A8xe2x80x94A9xe2x80x94,
(e) R1axe2x80x94A5xe2x80x94A6xe2x80x94A7xe2x80x94A8xe2x80x94A9xe2x80x94,
(f) R1axe2x80x94A6xe2x80x94A7xe2x80x94A8xe2x80x94A9xe2x80x94,
(g) R1axe2x80x94A7xe2x80x94A8xe2x80x94A9xe2x80x94,
(h) R1axe2x80x94A8xe2x80x94A9xe2x80x94,
(i) R1axe2x80x94A9xe2x80x94, and
(j) R1axe2x80x94;
Y is selected from the group consisting of
(a) xe2x80x94R3,
(b) xe2x80x94A28xe2x80x94R3,
(c) xe2x80x94A28xe2x80x94A29xe2x80x94R3,
(d) xe2x80x94A28xe2x80x94A29xe2x80x94A30xe2x80x94R3,
(e) xe2x80x94A28xe2x80x94A29xe2x80x94A30xe2x80x94A31xe2x80x94R3,
(f) xe2x80x94A28xe2x80x94A29xe2x80x94A30xe2x80x94A31xe2x80x94A32xe2x80x94R3,
(g) xe2x80x94A28xe2x80x94A29xe2x80x94A30xe2x80x94A31xe2x80x94A32xe2x80x94A33xe2x80x94R3, and
(h) xe2x80x94A28xe2x80x94A29xe2x80x94A30xe2x80x94A31xe2x80x94A32xe2x80x94A33xe2x80x94A34xe2x80x94R3;
R1a is H, alkyl, aralkyl or xe2x80x94COR2;
R1b is R1a or a group of formula 
R2 is alkyl, alkenyl, alkynyl, aryl or aralkyl;
R3 is a group of formula A35xe2x80x94OR4 or A35xe2x80x94NR4R5;
R4 and R5 are independently H or lower alkyl;
R6 and R9 are independently H or alkyl;
R7 is alkyl;
R8 is H, alkyl or COR2;
R10 is H or halogen;
R11 is alkyl or aralkyl;
A0 is absent or a peptide of from one to six amino acid residues;
A1 is Ser, Ala, Gly or D-Pro, or an equivalent amino acid thereof;
A2 is Ala, Val or Gly, or an equivalent amino acid thereof;
A3 is Ala, Ser, Gly or D-Pro, or an equivalent amino acid thereof;
A4 is Glu, Ala or Gly, or an equivalent amino acid thereof;
A5 is Ile, His, Ala or Gly, or an equivalent amino acid thereof;
A6 is Ala, Gln, Gly or D-Pro, or an equivalent amino acid thereof;
A7 is Ala, Leu or Gly, or an equivalent amino acid thereof;
A8 is Leu, Nle, Gly or D-Pro, or an equivalent amino acid thereof;
A9 is His, Ala, Gly or D-Pro, or an equivalent amino acid thereof;
A10 is Ala, Asn, Gly Lys, Asp or D-Pro, or an equivalent amino acid thereof;
A11 is Ala, Gly, Leu or Lys, or an equivalent amino acid thereof;
A12 is Ala or Gly, or an equivalent amino acid thereof;
A13 is Ala, Gly or Lys, or an equivalent amino acid thereof;
A14 is Ala, Gly, His, Ser, Asp, Lys or D-Pro, or an equivalent amino acid thereof;
A15 is Ala, Gly, Ile, D-Pro or Leu, or an equivalent amino acid thereof;
A16 is Asn, Ala, Gly, D-Pro or Gln, or an equivalent amino acid thereof;
A17 is Ala, Asp, Gly, Ser, Lys or D-Pro, or an equivalent amino acid thereof;
A18 is Lys, or an equivalent amino acid thereof;
A19 is Arg or Glu, or an equivalent amino acid thereof;
A20 is Arg, or an equivalent amino acid thereof;
A21 is Arg, Lys, Asp or Val, or an equivalent amino acid thereof;
A22 is Asp, Lys, Orn or Glu, or an equivalent amino acid thereof;
A23 is Leu, Phe or Trp, or an equivalent amino acid thereof;
A24 is Leu, or an equivalent amino acid thereof;
A25 is Arg, His, Asp, Lys or Glu, or an equivalent amino acid thereof;
A26 is Lys or His , or an equivalent amino acid thereof;
A27 is Leu or Lys, or an equivalent amino acid thereof;
A28 is Ile or Leu, or an equivalent amino acid thereof;
A29 is Ala, Asp, Glu or Gln, or an equivalent amino acid thereof;
A30 is Asp, Lys or Glu, or an equivalent amino acid thereof;
A31 is Ile, Leu or Val, or an equivalent amino acid thereof;
A32 is His , or an equivalent amino acid thereof;
A33 is Asn, or an equivalent amino acid thereof; and
A34 is Ala or Phe, or an equivalent amino acid thereof; and
A35 is absent or a peptide of from 1 to 4 amino acids.
The peptide compounds of the present invention possess useful properties, more particularly pharmaceutical properties. They are especially useful for treating disease states capable of being modulated by compounds which bind to parathyroid hormone receptors either with or without comcommitant stimulation of adenylyl cyclase activity. The present invention is therefore also directed to the pharmaceutical use of the peptide compounds and pharmaceutical compositions containing the peptide compounds.
As used above and throughout the specification, the following terms, unless otherwise indicated, shall be understood to have the following meanings.
xe2x80x9cPatientxe2x80x9d includes both human and other mammals.
xe2x80x9cAlkylxe2x80x9d means an aliphatic hydrocarbon group which may be straight or branched having about 1 to about 20 carbon atoms in the chain. Branched means that one or more lower alkyl groups are attached to a linear alkyl chain. xe2x80x9cLower alkylxe2x80x9d means about 1 to 4 carbon atoms in the chain which may be straight or branched. Alkyl groups are exemplified by methyl, ethyl, n- and iso-propyl, n-, sec-, iso- and tert-butyl, and the like.
xe2x80x9cAlkenylxe2x80x9d means aliphatic hydrocarbon group containing a carbon-carbon double bond and which may be straight or branched having about 2 to about 20 carbon atoms in the chain. xe2x80x9cLower alkenylxe2x80x9d means about 2 to 4 carbon atoms in the chain which may be straight or branched. Exemplary alkenyl groups include ethenyl, propenyl, n-butenyl, i-butenyl, 3-methylbut-2-enyl, n-pentenyl, heptenyl, octenyl, cyclohexylbutenyl and decenyl.
xe2x80x9cAlkynylxe2x80x9d means aliphatic hydrocarbon group containing a carbon-carbon triple bond and which may be straight or branched having about 2 to about 20 carbon atoms in the chain. xe2x80x9cLower alkynylxe2x80x9d means about 2 to 4 carbon atoms in the chain which may be straight or branched. Exemplary alkynyl groups include ethynyl, propynyl, n-butynyl, 3-methylbut-2-ynyl, n-pentynyl, heptynyl, octynyl and decynyl.
xe2x80x9cAlkylenexe2x80x9d denotes a divalent group derived from a straight or branched chain saturated hydrocarbon by the removal of two hydrogen atoms, for example methylene, 1,2-ethylene, 1,1-ethylene, 1,3-propylene, 2,2-dimethylpropylene, and the like.
xe2x80x9cAralkylxe2x80x9d means an aryl group attached to the parent molecular moiety through an alkylene. Preferred aralkyls contain a lower alkyl moiety. Representative aralkyl groups include benzyl, 2-phenethyl, naphthlenemethyl. and the like. A preferred aralkyl group is benzyl.
xe2x80x9cArylxe2x80x9d means an aromatic monocyclic or multicyclic ring system of 6 to about 14 carbon atoms. preferably of about 6 to about 10 carbon atoms. The aryl is optionally substituted with one or more substituents selected from alkyl, hydroxy, halogen and haloalkyl. Representative aryl groups include phenyl and naphtlyl.
xe2x80x9cAmino acidxe2x80x9d means an amino acid selected from the group consisting of natural and unnaturai amino acids as defined herein. The amino acids may be neutral, positive or negative depending on the substituents in the side chain. xe2x80x9cNeutral amino acidxe2x80x9d means an amino acid containing uncharged side chain substituents. Exemplary neutral amino acids include alanine, valine, leucine, isoleucine, proline, phenylalanine, tryptophan, methionine, glycine, serine, threonine and cysteine. xe2x80x9cPositive amino acidxe2x80x9d means an amino acid in which the side chain substituents are positively charged at physiological pH. Exemplary positive amino acids include lysine, arginine and histidine. xe2x80x9cNegative amino acidxe2x80x9d means an amino acid in which the side chain substituents bear a net negative charge at physiological pH. Exemplary negative amino acids include aspartic acid and glutamic acid. Preferred amino acids are xcex1-amino acids. The most preferred amino acids are xcex1-amino acids having L stereochemistry at the xcex1-carbon.
xe2x80x9cNatural amino acidxe2x80x9d means an xcex1-amino acid selected from the group consisting of alanine, valine, leucine, isoleucine, proline, phenylalanine, tryptophan, methionine, glycine, serine, threonine, cysteine, tyrosine, asparagine, glutamine, lysine, arginine, histidine, aspartic acid and glutamic acid.
xe2x80x9cUnnatural amino acidxe2x80x9d means an amino acid for which is no nucleic acid codon. Examples of unnatural amino acids include, for example, the D-isomers of the natural xcex1-amino acids such as D-proline (D-P, D-Pro) as indicated above; Aib (aminobutyric acid), bAib (3-aminoisobutyric acid), Nva (norvaline), xcex2-Ala, Aad (2-aminoadipic acid), bAad (3-aminoadipic acid), Abu (2-aminobutyric acid), Gaba (xcex3-aminobutyric acid), Acp (6-aminocaproic acid), Dbu (2,4-diaminobutryic acid), xcex1-aminopimelic acid, TMSA (trimethylsilyl-Ala), aIle (allo-isoleucine), Nle (norleucine), tert-Leu, Cit (citrulline), Orn (ornithine, O), Dpm (2,2xe2x80x2-diaminopimelic acid), Dpr (2,3-diaminopropionic acid), xcex1- or xcex2-Nal, Cha (cyclohexyl-Ala), hydroxyproline, Sar (sarcosine), and the like; cyclic amino acids; Nxcex1-alkylated amino acids such as MeGly (Nxcex1-methylglycine), EtGly (Nxcex1-ethylglycine) and EtAsn (Nxcex1-ethylasparagine); and amino acids in which the xcex1-carbon bears two side-chain substituents.
xe2x80x9cPeptidexe2x80x9d and xe2x80x9cpolypeptidexe2x80x9d mean a polymer in which the monomers are amino acid residues joined together through amide bonds. Preferred peptide compounds of the present invention are those comprising xcex1-amino acids. xe2x80x9cPeptide compoundxe2x80x9d means a compound comprising a peptide as defined herein.
xe2x80x9cAmino acid residuexe2x80x9d means the individual amino acid units incorporated into the peptide compounds of the invention.
The names of natural and unnatural amino acids and residues thereof used herein follow the naming conventions suggested by the IUPAC Commission on the Nomenclature of Organic Chemistry and the IUPAC-IUB Commission on Biochemical Nomenclature as set out in xe2x80x9cNomenclature of xcex1-Amino Acids (Recommendations, 1974)xe2x80x9d Biochemistry, 14(2), (1975). To the extent that the names and abbreviations of amino acids and residues thereof employed in this specification and appended claims differ from those noted, differing names and abbreviations will be made clear.
xe2x80x9cEquivalent amino acidxe2x80x9d means an amino acid which may be substituted for another amino acid in the peptide compounds according to the invention without any appreciable loss of function. In making such changes, substitutions of like amino acids is made on the basis of relative similarity of side chain substituents, for example regarding size, charge, hydrophilicity, hydropathicity and hydrophobicity as described herein. The phrase xe2x80x9cor an equivalent amino acid thereofxe2x80x9d when used following a list of individual amino acids means an equivalent of each of the individual amino acids included in the list.
As detailed in U.S. Pat. No. 4,554,101, incorporated herein by reference, the following hydrophilicity values have been assigned to amino acid residues: Arg (+3.0); Lys (+3.0); Asp (+3.0); Glu (+3.0); Ser (+0.3); Asn (+0.2); Gln (+0.2); Gly (0); Pro (xe2x88x920.5); Thr (xe2x88x920.4); Ala (xe2x88x920.5); Cys (xe2x88x921.0); Met (xe2x88x921.3); Val (xe2x88x921.5); Leu (xe2x88x921.8); Ile (xe2x88x921.8); Tyr (xe2x88x922.3); Phe (xe2x88x922.5); and Trp (xe2x88x923.4). It is understood that an amino acid residue can be substituted for another having a similar hydrophilicity value (e.g., within a value of plus or minus 2.0) and still obtain a biologically equivalent polypeptide.
In a similar manner, substitutions can be made on the basis of similarity in hydropathic index. Each amino acid residue has been assigned a hydropathic index on the basis of its hydrophobicity and charge characteristics. Those hydropathic index values are: Ile (+4.5); Val (+4.2); Leu (+3.8); Phe (+2.8); Cys (+2.5); Met (+1.9); Ala (+1.8); Gly (xe2x88x920.4); Thr (xe2x88x920.7); Ser (xe2x88x920.8); Trp (xe2x88x920.9); Tyr (xe2x88x921.3); Pro (xe2x88x921.6); His (xe2x88x923.2); Glu (xe2x88x923.5); Gln (xe2x88x923.5); Asp (xe2x88x923.5); Asn (xe2x88x923.5); Lys (xe2x88x923.9); and Arg (xe2x88x924.5). In making a substitution based on the hydropathic index, a value of within plus or minus 2.0 is preferred.
In the peptide compounds of this invention, the ester, amide, disulfide or lanthionine bond which links two amino acid residues is formed between the side-chain functionalities. Thus, an amide is bond is formed between the side-chain carboxyl group of an acidic amino acid residue and the side-chain amino group of a basic amino acid residue. Preferred acidic amino acid residues include Asp, Glu, xe2x80x94NHCH[(CH2)3CO2H]COxe2x80x94 and xe2x80x94NHCH[(CH2)4CO2H]COxe2x80x94, Asp being most preferred. Preferred basic amino acid residues include His, Lys, Orn, xe2x80x94NHCH(CH2NH2)COxe2x80x94 and xe2x80x94NHCH[(CH2)2NH2]COxe2x80x94, Lys being most preferred.
Ester bonds are formed between the side-chain carboxyl group of an acidic amino acid residue as described above and the side chain hydroxy group of an amino acid residue such as Ser, Thr, Tyr and the like, Ser and Thr being especially preferred.
Disulfides are formed from amino acid residues containing side chain sulfhydryl groups. Cys is especiaiiy preferred for the tormation or disulfide bonds. Lanthionine bridges are formed by desulfurization of the corresponding disulfide.
The number of atoms in the bridge resulting from the amide, ester, disulfide or lanthionine bond formed as described above will vary depending on the length of the side chain and the type of bond (i.e., amide, ester, disulfide or lanthionine). The bridge preferably comprises from 4 to 12 atoms, more preferably from 6 to 10 atoms. A further preferred number of atoms contained in the bridge is 7, this bridge preferably comprising an amide bond between the side-chain functionalities of a Lys and an Asp residue.
A representative peptide compound of the present invention is denoted, for example, as cyclo(K18xe2x80x94D22)[A1, Nle8, K18, D22, L27]hPTH(1-31)NH2 with the linked amino acid residues in the parenthesis following xe2x80x9ccycloxe2x80x9d and substituted amino acids from the natural sequence are placed in brackets. hPTH stands for human parathyroid hormone and hPTHrP for human parathyroid hormone-related protein. The numbers in the second parenthesis refer to the number of amino acid residues in the peptide compound, beginning at the N-terminus (i.e., the first 31 amino acids of hPTH).
Where the peptide compound of the present invention is substituted with a basic moiety, acid addition salts are formed and are simply a more convenient form for use; and in practice, use of the salt form inherently amounts to use of the free base form. The acids which can be used to prepare the acid addition salts include preferably those which produce, when combined with the free base, pharmaceutically acceptable salts, that is, salts whose anions are non-toxic to the patient in pharmaceutical doses of the salts, so that the beneficial effects inherent in the free base are not vitiated by side effects ascribable to the anions. Although pharmaceutically acceptable salts of said basic compounds are preferred, all acid addition salts are useful as sources of the free base form even if the particular salt, per se, is desired only as an intermediate product as, for example, when the salt is formed only for purposes of purification, and identification, or when it is used as intermediate in preparing a pharmaceutically acceptable salt by ion exchange procedures. Pharmaceutically acceptable salts within the scope of the invention are those derived from the following acids: mineral acids such as hydrochloric acid, sulfuric acid, phosphoric acid and sulfamic acid; and organic acids such as acetic acid, citric acid, lactic acid, tartaric acid, malonic acid, methanesufoniic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, cyclohexylsulfamic acid, quinic acid, and the like. The corresponding acid addition salts comprise the following: hydrohalides, e.g. hydrochloride and hydrobromide, sulfate, phosphate, nitrate, sulfamate, acetate, citrate, lactate, tartarate, malonate, oxalate, salicylate, propionate, succinate, fumarate, maleate, methylene-bis-xcex2-hydroxynaphthoates, gentisates, mesylates, isethionates and di-p-toluoyltartratesmethanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, cyclolhexylsulfamate and quinate, respectively.
According to a further feature of the invention, acid addition saits of the peptide compounds of this invention are prepared by reaction of the free base with the appropriate acid, by the application or adaptation of known methods. For example, the acid addition salts of the peptide compounds of this invention are prepared either by dissolving the free base in aqueous or aqueous-alcohol solution or other suitable solvents containing the appropriate acid and isolating the salt by evaporating the solution, or by reacting the free base and acid in an organic solvent, in which case the salt separates directly or can be obtained by concentration of the solution.
Preferred acid addition salts are the trifluoroacetate, acetate and hydrochloride. The acetate and tetrahydrochloride salts are especially preferred.
The peptide compounds of this invention can be regenerated from the salts by the application or adaptation of known methods. For example, parent peptide compounds of the invention can be regenerated from their acid addition salts by treatment with an alkali, e.g. aqueous sodium bicarbonate solution or aqueous ammonia solution.
Where the peptide compound of the invention is substituted with an acidic moiety, base addition salts may be formed and are simply a more convenient form for use; and in practice, use of the salt form inherently amounts to use of the free acid form. The bases which can be used to prepare the base addition salts include preferably those which produce, when combined with the free acid, pharmaceutically acceptable salts, that is, salts whose cations are non-toxic to the animal organism in pharmaceutical doses of the salts, so that the beneficial effects inherent in the free acid are not vitiated by side effects ascribable to the cations. Pharmaceutically acceptable salts, including for example alkali and alkaline earth metal salts, within the scope of the invention are those derived from the following bases: sodium hydride, sodium hydroxide, potassium hydroxide, calcium hydroxide, aluminum hydroxide, lithium hydroxide, magnesium hydroxide, zinc hydroxide, ammonia, trimethylammonia, triethylammonia, ethylenediamine, n-methyl-glucamine, lysine, arginine, ornithine, choline, N,Nxe2x80x2-dibenzylethylenediamine, chloroprocaine, diethanolamine, procaine, n-benzylphenethylamine, diethylamine, piperazine, tris(hydroxymethyl)aminomethane, tetramethylammmonium hydroxide, and the like.
Metal salts of peptide compounds of the present invention may be obtained by contacting a hydride, hydroxide, carbonate or similar reactive compound of the chosen metal in an aqueous or organic solvent with the free acid form of the peptide compound. The aqueous solvent employed may be water or it may be a mixture of water with an organic solvent, preferably an alcohol such as methanol or ethanol, a ketone such as acetone, an aliphatic ether such as tetrahydrofuran, or an ester such as ethyl acetate. Such reactions are normally conducted at ambient temperature but they may, if desired, be conducted with heating.
Amine salts of peptide compounds of the present invention may be obtained by contacting an amine in an aqueous or organic solvent with the free acid form of the peptide compound. Suitable aqueous solvents include water and mixtures of water with alcohols such as methanol or ethanol, ethers such as tetrahydrofuran, nitriles such as acetonitrile, or ketones such as acetone. Amino acid salts may be similarly prepared.
The base addition salts of the peptide compounds of this invention can be regenerated from the salts by the application or adaptation of known methods. For example, parent peptide compounds of the invention can be regenerated from their base addition salts by treatment with an acid, e.g. hydrochloric acid.
As well as being useful in themselves as active compounds, salts of peptide compounds of the invention are useful for the purposes of purification of the peptide compounds, for example by exploitation of the solubility differences between the salts and the parent peptide compounds. side products and/or starting materials by techniques well known to those skilled in the art.
xe2x80x9cPharmaceutically acceptable esterxe2x80x9d means esters which hydrolyze in vivo and include those that break down readily in the human body to leave the parent peptide compound or a salt thereof. Suitable ester groups include, for example, those derived from pharmaceutically acceptable aliphatic carboxylic acids, particularly alkanoic, alkenoic, cycloalkanoic and alkanedioic acids, in which each alkyl or alkenyl moiety advantageously has not more than 6 carbon atoms. Examples of particular esters includes formates, acetates, propionates, butyrates, acrylates and ethylsuccinates.
xe2x80x9cProdrugxe2x80x9d means a compound which is rapidly transformed in vivo to yield the parent peptide compound, for example by hydrolysis in blood. xe2x80x9cPharmaceutically acceptable prodrugxe2x80x9d means a compound which is, within the scope of sound medical judgement, suitable for pharmaceutical use in a patient without undue toxicity, irritation, allergic response, and the like, and effective for the intended use, including a pharmaceutically acceptable ester as well as a zwitterionic form, where possible, of the peptide compounds of the invention. Pharmaceutically acceptable prodrugs according to the invention are described in T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems, Vol. 14 of the A.C.S. Symposium Series, and in Edward B. Roche, ed., Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press, 1987, both of which are incorporated herein by reference.
xe2x80x9cSolvatexe2x80x9d means a physical association of a compound of this invention with one or more solvent molecules. This physical association involves varying degrees of ionic and covalent bonding, including hydrogen bonding. In certain instances the solvate will be capable of isolation, for example when one or more solvent molecules are incorporated in the crystal lattice of the crystalline solid. xe2x80x9cSolvatexe2x80x9d encompasses both solution-phase and isolable solvates. Representative solvates include ethanoiates, methanolates, and the like. xe2x80x9cHydratexe2x80x9d is a solvate wherein the solvent molecule(s) is/are H2O.
The peptide compounds of the present invention may contain asymmetric centers in addition to the chiral centers in the backbone of the peptide compound. These asymmetric centers may independently be in either the R or S configuration. It will also be apparent to those skilled in the art that certain peptide compounds of formula I may exhibit geometrical isomerism. Geometrical isomers include the cis and trans forms of peptide compounds of the invention having alkenyl moieties. The present invention comprises the individual geometrical isomers and stereoisomers and mixtures thereof.
Such isomers can be separated from their mixtures. by the application or adaptation of known methods, for example chromatographic techniques and recrystallization techniques, or they are separately prepared from the appropriate isomers of their intermediates, for example by the application or adaptation of methods described herein.
Peptide compounds contemplated as falling within the scope of the present invention include, but are not limited to
cyclo(K18xe2x80x94D22)[A1,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 3)
[A1,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 4)
cyclo(K18xe2x80x94D22)[A1,2Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 5)
cyclo(K18xe2x80x94D22)[A1,3Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 6)
cyclo(K18xe2x80x94D22)[A1,4Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 7)
cyclo(K18xe2x80x94D22)[A1,5Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 8)
cyclo(K18xe2x80x94D22)[A1,6Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 9)
cyclo(K18xe2x80x94D22)[A1,7Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 10)
cyclo(K18xe2x80x94D22)[A1,9Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 11)
cyclo(K18xe2x80x94D22)[A1,10Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 12)
cyclo(K18xe2x80x94D22)[A1,11Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 13)
cyclo(K18xe2x80x94D22)[A1,12Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 14)
cyclo(K18xe2x80x94D22)[A1,13Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 15)
cyclo(K18xe2x80x94D22)[A1,14Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 16)
cyclo(K18xe2x80x94D22)[A1,15Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 17)
cyclo(K18xe2x80x94D22)[A1,16Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 18)
cyclo(K18xe2x80x94D22)[A1,17Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 19)
cyclo(K18xe2x80x94D22)[G1,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 20)
cyclo(K18xe2x80x94D22)[A1,G2Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 21)
cyclo(K18xe2x80x94D22)[A1,G3Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 22)
cyclo(K18xe2x80x94D22)[A1,G4Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 23)
cyclo(K18xe2x80x94D22)[A1,G5Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 24)
cyclo(K18xe2x80x94D22)[A1,G6Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 25)
cyclo(K18xe2x80x94D22)[A1,G7Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 26)
cyclo(K18xe2x80x94D22)[A1,G8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 27)
cyclo(K18xe2x80x94D22)[A1,G9Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 28)
cyclo(K18xe2x80x94D22)[A1,G10Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 29)
cyclo(K18xe2x80x94D22)[A1,G11Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 30)
cyclo(K18xe2x80x94D22)[A1,G13Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 31)
cyclo(K18xe2x80x94D22)[A1,G14Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 32)
cyclo(K18xe2x80x94D22)[A1,G15Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 33)
cyclo(K18xe2x80x94D22)[A1,G16Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 34)
cyclo(K18xe2x80x94D22)[A1,G17Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 35)
cyclo(K18xe2x80x94D22)[Dxe2x80x94P1,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 36)
cyclo(K18xe2x80x94D22)[A1,Dxe2x80x94P3,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 37)
cyclo(K18xe2x80x94D22)[A1,Dxe2x80x94P6,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 38)
cyclo(K18xe2x80x94D22)[A1,Dxe2x80x94P7,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 39)
cyclo(K18xe2x80x94D22)[A1,Dxe2x80x94P9,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 40)
cyclo(K18xe2x80x94D22)[A1,Dxe2x80x94P10,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 41)
cyclo(K18xe2x80x94D22)[A1,Dxe2x80x94P14,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 42)
cyclo(K18xe2x80x94D22)[A1,Dxe2x80x94P15,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 43)
cyclo(K18xe2x80x94D22)[A1,Dxe2x80x94P16,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 44)
cyclo(K18xe2x80x94D22)[A1,Dxe2x80x94P17,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 45)
cyclo(K18xe2x80x94D22)[A1,Nle8,K18,D22,L27]hPTH(1-34)NH2 (SEQ ID NO: 46)
cyclo(D18xe2x80x94K22)[A1,Nle8,D18,K22,L27]hPTH(1-31)NH2 (SEQ ID NO: 47)
cyclo(O18xe2x80x94D22)[A1,Nle8,O18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 48)
cyclo(D18xe2x80x94O22)[A1,Nle8,D18,O22,L27]hPTH(1-31)NH2 (SEQ ID NO: 49)
cyclo(K18xe2x80x94E22)[A1,Nle8,K18,E22,L27]hPTH(1-31)NH2 (SEQ ID NO: 50)
cyclo(O18xe2x80x94E22)[A1,Nle8,O18,E22,L27]hPTH(1-31)NH2 (SEQ ID NO: 51)
cyclo(K18xe2x80x94D22)[A1,Nle8,K18,D22,L27]hPTH(1-30)NH2 (SEQ ID NO: 52)
cyclo(K18xe2x80x94D22)[A1,Nle8,K18,D22,L27]hPTH(1-29)NH2 (SEQ ID NO: 53)
cyclo(K18xe2x80x94D22)[A1,Nle8,K18,D22,L27]hPTH(1-28)NH2 (SEQ ID NO: 54)
cyclo(K18xe2x80x94D22)[A1,Nle8,K18,D22,L27]hPTH(1-27)NH2 (SEQ ID NO: 55)
cyclo(K18xe2x80x94D22)[K18,D22,L27]hPTH(10-31)NH2 (SEQ ID NO: 56)
cyclo(K18xe2x80x94D22)[K18,D22,L27]hPTH(9-31)NH2 (SEQ ID NO: 57)
cyclo(K18xe2x80x94D22)[Nle8,K18,D22,L27]hPTH(8-31)NH2 (SEQ ID NO: 58)
cyclo(K18xe2x80x94D22)[Nle8,K18,D22,L27]hPTH(7-31)NH2 (SEQ ID NO: 59)
cyclo(K18xe2x80x94D22)[Nle8,K18,D22,L27]hPTH(6-31)NH2 (SEQ ID NO: 60)
cyclo(K18xe2x80x94D22)[Nle8,K18,D22,L27]hPTH(5-31)NH2 (SEQ ID NO: 61)
cyclo(K18xe2x80x94D22)[Nle8,K18,D22,L27]hPTH(4-31)NH2 (SEQ ID NO: 62)
cyclo(K18xe2x80x94D22)[Nle8,K18,D22,L27]hPTH(3-31)NH2 (SEQ ID NO: 63)
cyclo(K18xe2x80x94D22)[Nle8,K18,D22,L27]hPTH(2-31)NH2 (SEQ ID NO: 64)
cyclo(K18xe2x80x94D22)[Nle8,K18,D22,L27]hPTH(7-34)NH2 (SEQ ID NO: 65)
cyclo(K10xe2x80x94D14)[A1,Nle8,18,K10,D14,L27]hPTH(1-31)NH2 (SEQ ID NO: 66)
cyclo(K14xe2x80x94D18)[A1,Nle8,K14,D18,L27]hPTH(1-31)NH2 (SEQ ID NO: 67)
cyclo(K17xe2x80x94D21)[A1,Nle8,18,K17,D21,L27]hPTH(1-31)NH2 (SEQ ID NO: 68)
cyclo(K21xe2x80x94D25)[A1,Nle8,18,K21,D25,L27]hPTH(1-31)NH2 (SEQ ID NO: 69)
cyclo(K25xe2x80x94D29)[A1,Nle8,18,K25,D29,L27]hPTH(1-31)NH2 (SEQ ID NO: 70)
cyclo(K18xe2x80x94D22)[K18,D22,L27]hPTH(1-34)NH2 (SEQ ID NO: 71)
cyclo(K18xe2x80x94D22)[K18,26,30,D22,L23,28,31,E25,29]hPTH(1-34)NH2 (SEQ ID NO: 72)
bicyclo(K13xe2x80x94D17,K18xe2x80x94D22)[A1,Nle8,D17,22,L27]hPTH(1-31)NH2 (SEQ ID NO: 73)
bicyclo(K18xe2x80x94D22,K26xe2x80x94D30)[A1,Nle8,K18D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 74)
bicyclo(K13xe2x80x94D17,K18xe2x80x94D22)[A1,Nle8,D17,22,K18,L27]hPTH(1-34)NH2 (SEQ ID NO: 75)
cyclo(K18xe2x80x94D22)[K18,D22]hPTH(7-34)NH2 (SEQ ID NO: 77)
bicyclo(K13xe2x80x94D17,K18xe2x80x94D22)[Nle8,K,D17,22,L27]hPTH(7-34)NH2 (SEQ ID NO: 78)
bicyclo(K18xe2x80x94D22,K26xe2x80x94D30)[Nle8,K18,D22,L27]hPTH(7-34)NH2 (SEQ ID NO: 79)
tricyclo(K13xe2x80x94D17,K18xe2x80x94D22,K26xe2x80x94D30)[A1,Nle8,K18,D17,22,L27]hPTH(1-31)NH2 (SEQ ID NO: 80)
or a pharmaceutically acceptable salt or prodrug thereof.
A preferred cyclic peptide compound of formula 2 has formula I above wherein the bridge formed from the side chains of one pair of amino acid residues is non-overlapping with a bridge formed between the side chains of another pair of amino acid residues.
A more preferred cyclic peptide compound of formula 3 has formula 2 above wherein A10 is Ala, Asn, Asp, Gly or Lys; A13 is Ala, Gly or Lys; A14 is Ala, Asp, Gly, His, Lys or Ser; A17 is Ala, Asp, Gly, Lys or Ser; A18 is Asp, Leu, Lys, Orn or Nle; A2, is Arg, Asp, Lys or Val; A22 is Asp, Glu, Lys, Orn or Phe; A25 is Arg, Asp, Glu, His or Lys; A26 is His or Lys; A29 is Ala, Asp, Glu or Gln; A30 is Asp, Glu or Lys, and
the side chains of at least one of the following pairs of amino acid residues, A10 and A14, A13 and A17, A14 and A18, A17 and A21, A18 and A22, A21 and A25, A25 and A29 and A26 and A30 are linked through an amide bond to form a bridge, and the side chain of each of the following amino acid residues, A10, A13, A14, A17, A18, A21, A22, A25, A26, A29, and A30, contributes, at most, to the formation of a single and non-overlapping bridge; provided
(a) that when the side chains of the pair of amino acid residues A13 and A17 are linked through an amide bond to form a bridge, then the side chains of at least one of the following pairs of amino acid residues, A18 and A22, A21 and A25, and A25 and A29 are also linked through an amide bond to form a bridge;
(b) that when the side chains of the following pair of amino acid residues A26 and A30 are linked through an amide bond to form a bridge, then the side chains of at least one of the following pairs of amino acid residues A10 and A14, A14 and A18, A17 and A21, A18 and A22 and A21 and A25 are also linked through an amide bond to form a bridge; and
(c) that when the side chains of the following pairs of amino acid residues A13 and A17 and A26 and A30 are linked through an amide bond to form a bridge, then the side chains of one of the following pairs of amino acid residues A18 and A22 and A21 and A25 are also linked through an amide bond to form a bridge.
Another more preferred cyclic peptide compound of formula 4 has formula 3 above wherein R1a is H and Y is NH2.
Certain cyclic peptide compounds of this invention possess agonist activity on the parathyroid hormone receptor and accordingly are useful in the treatment of physiological conditions associated with bone cell calcium regulation including hypocalcemia; osteoporosis; osteopenia; and disorders associated with osteoporosis and osteopenia such as hyperparathyroidism, hypoparathyroidism, and Cushings syndrome; glucocorticoid- and immunosuppressant-induced osteopaenia: and bone fracture and bone refracture repair.
A preferred cyclic peptide agonist compound of formula 5 has formula 4 above wherein X is
(a) R1axe2x80x94A1xe2x80x94A2xe2x80x94A3xe2x80x94A4xe2x80x94A5xe2x80x94A6xe2x80x94A7xe2x80x94A8xe2x80x94A9xe2x80x94,
(b) R1axe2x80x94A2xe2x80x94A3xe2x80x94A4xe2x80x94A5xe2x80x94A6xe2x80x94A7xe2x80x94A8xe2x80x94A9xe2x80x94 or
(c) R1axe2x80x94A3xe2x80x94A4xe2x80x94A5xe2x80x94A6xe2x80x94A7xe2x80x94A8xe2x80x94A9xe2x80x94.
A more preferred cyclic peptide agonist compound of formula 6 has formula 5 above wherein A1 is Ala, Gly or D-Pro; A1 is Nle and A27 is Leu.
Another more preferred cyclic peptide agonist compound of formula 7 has formula 6 above wherein
(i) the side chains of A10 and A14 are linked through an amide bond to form a bridge;
(ii) the side chains of A14 and A18 are linked through an amide bond to form a bridge;
(iii) the side chains of A17 and A21 are linked through an amide bond to form a bridge;
(iv) the side chains of A18 and A22 are linked through an amid bond to form a bridge;
(v) the side chains of A21 and A25 are linked through an amide bond to form a bridge; or
(vi) the side chains of A25 and A29 are linked through an amide bond to form a bridge.
Another more preferred cyclic peptide agonist compound has formula (i) above wherein A10 is Asp or Lys; A13 is Lys; A14 is Asp or Lys; A17 is Asp or Ser; A18 is Nle; A21 is Arg or Val; A22 is Glu or Phe; A25 is Arg or His; A26 is Lys or His, A29 is Ala or Gln; and A30 is Asp or Glu; and the side-chains of A10 and A14 are linked through an amide bond to form a bridge.
Another more preferred cyclic peptide agonist compound has formula (ii) above wherein A10 is Asn or Asp; A13 is Lys; A14 is Asp or Lys; A17 is Asp or Ser; A18 is Nle; A21 is Arg or Val; A22 is Glu or Phe; A25 is Arg or His; A26 is His or Lys; A29 is Ala or Gln; and A30 is Asp or Glu; and the side chains of A14 and A18 are linked through an amide bond to form a bridge.
Another more preferred cyclic peptide agonist compound has formula (iii) above wherein A10 is Asn or Asp; A13 is Lys; A14 is His or Ser; A17 is Asp or Lys; A18 is Nle; A21 is Asp or Lys; A21 is Glu or Phe; A25 is Arg or His; A26 is His or Lys; A29 is Ala or Gln; and A30 is Asp or Glu; and the side chains of A17 and A21 are linked through an amide bond to form a bridge.
Another more preferred cyclic peptide agonist compound has formula (iv) above wherein A10 is Asn or Asp; A13 is Lys; A14 is His or Ser; A17 is Asp or Ser; A18 is Asp, Lys or Orn; A21 is Arg or Val; A22 is Asp, Glu, Lys or Orn; A25 is Arg or His; A26 is His or Lys; A29 is Ala or Gln; and A30 is Asp or Glu; and the side chains of A18 and A22 are linked through an amide bond to form a bridge.
Another more preferred cyclic peptide agonist compound has formula (v) above wherein A10 is Asn or Asp; A13 is Lys; A14 is His or Ser; A17 is Asp or Ser; A18 is Nle; A21 is Asp or Lys; A22 is Glu or Phe; A25 is Asp or Lys; A26 is His or Lys; A29 is Ala or Gln; and A30 is Asp or Glu; and the side chains of A21 and A25 are linked through an amide bond to form a bridge.
Another more preferred cyclic peptide agonist compound has formula (vi) above wherein A10 is Asn or Asp; A13 is Lys; A14 is His or Ser; A17 is Asp or Ser; A18 is Nle; A21 is Arg or Val; A22 is Glu or Phe; A25 is Asp or Lys; A26 is His or Lys; A29 is Asp or Lys; and A30 is Asp or Glu; and the side chains of A25 and A29 are linked through an amide bond to form a bridge.
Another more preferred cyclic peptide agonist compound of formula 8 has formula 6 above wherein (vii) the side-chains of A13 and A17 are linked through an amide bond and the side-chains of A18 and A22 are linked through an amide bond to form a bridge; or (viii) the side-chains of A18 and A22 are linked through an amide bond and the side-chains of A26 and A30 are linked through an amide bond to form a bridge.
Another more preferred cyclic peptide agonist compound has formula (vii) above wherein A10 is Asn or Asp; A13 is Lys or Asp; A14 is His or Ser; A17 is Lys or Asp; A18 is Lys or Asp; A21 is Val or Arg; A22 is Glu, Lys or Asp; A25 is Arg or His; A26 is His or Lys; A29 is Ala or Gln; and A30 is Asp or Glu; and the side-chains of A13 and A17 are linked through an amide bond and the side-chains of A18 and A22 are linked through an amide bond to form a bridge.
Another more preferred cyclic peptide agonist compound has formula (viii) above wherein A10 is Asn or Asp; A13 is Lys; A14 is His or Ser; A18 is Ser or Asp; A18 is Lys or Asp; A21 is Val or Arg; A22 is Glu, Lys or Asp; A25 is Arg or His; A26 is Lys or Asp; A29 is Ala or Gln; and A30 is Lys or Asp; and the side-chains of A26 and A30 are linked through an amide bond and the side-chains of A18 and A22 are linked through an amide bond to form a bridge.
Another more preferred cyclic peptide agonist compound of formula 9 has formula 6 above wherein the side-chains of A13 and A17 are linked through an amide bond and the side-chains of A18 and A22 are linked through an amide bond and the side chains of A26 and A30 are linked through an amide bond to form a bridge.
Another more preferred cyclic peptide agonist compound of formula 10 has formula 9 above wherein A10 is Asn or Asp; A13 is Lys or Asp; A14 is His or Ser; A17 is Lys or Asp; A18 is Lys or Asp; A21 Val or Arg; A22 is Glu, Lys, or Asp, A25 is Arg or His; A26 is Lys or Asp; A29 is Ala or Gln; and A30 is Lys or Asp.
More preferred cyclic peptide agonist compounds of this invention include:
Cyclo(K18xe2x80x94D22)[A1,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 3);
Cyclo(K18xe2x80x94D22)[A1,2,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 5);
Cyclo(K18xe2x80x94D22)[A1,3,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 6);
Cyclo(K18xe2x80x94D22)[A1,4,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 7);
Cyclo(K18xe2x80x94D22)[A1,5,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 8);
Cyclo(K18xe2x80x94D22)[A1,6,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 9);
Cyclo(K18xe2x80x94D22)[A1,7,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 10);
Cyclo(K18xe2x80x94D22)[A1,9,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 11);
Cyclo(K18xe2x80x94D22)[A1,10,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 12);
Cyclo(K18xe2x80x94D22)[A1,11,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 13);
Cyclo(K18xe2x80x94D22)[A1,12,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 14);
Cyclo(K18xe2x80x94D22)[A1,13,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 15);
Cyclo(K18xe2x80x94D22)[A1,14,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 16);
Cyclo(K18xe2x80x94D22)[A1,15,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 17);
Cyclo(K18xe2x80x94D22)[A1,16,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 18);
Cyclo(K18xe2x80x94D22)[A1,17,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 19);
Cyclo(K18xe2x80x94D22)[G1,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 20);
Cyclo(K18xe2x80x94D22)[A1,G2,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 21);
Cyclo(K18xe2x80x94D22)[A1,G3,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 22);
Cyclo(K18xe2x80x94D22)[A1,G4,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 23);
Cyclo(K18xe2x80x94D22)[A1,G5,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 24);
Cyclo(K18xe2x80x94D22)[A1,G6,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 25);
Cyclo(K18xe2x80x94D22)[A1,G7,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 26);
Cyclo(K18xe2x80x94D22)[A1,G8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 27);
Cyclo(K18xe2x80x94D22)[A1,G9,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 28);
Cyclo(K18xe2x80x94D22)[A1,G10,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 29);
Cyclo(K18xe2x80x94D22)[A1,G11,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 30);
Cyclo(K18xe2x80x94D22)[A1,G13,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 31);
Cyclo(K18xe2x80x94D22)[A1,G14,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 32);
Cyclo(K18xe2x80x94D22)[A1,G15,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 33);
Cyclo(K18xe2x80x94D22)[A1,G16,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 34);
Cyclo(K18xe2x80x94D22)[A1,G17,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 35);
Cyclo(K18xe2x80x94D22)[Dxe2x80x94P1,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 36);
Cyclo(K18xe2x80x94D22)[A1,Dxe2x80x94P3,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 37);
Cyclo(K18xe2x80x94D22)[A1,Dxe2x80x94P6,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 38);
Cyclo(K18xe2x80x94D22)[A1,Dxe2x80x94P7,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 39);
Cyclo(K18xe2x80x94D22)[A1,Dxe2x80x94P9,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 40);
Cyclo(K18xe2x80x94D22)[A1,Dxe2x80x94P10,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 41);
Cyclo(K18xe2x80x94D22)[A1,Dxe2x80x94P14,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 42);
Cyclo(K18xe2x80x94D22)[A1,Dxe2x80x94P15,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 43);
Cyclo(K18xe2x80x94D22)[A1,Dxe2x80x94P16,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 44);
Cyclo(K18xe2x80x94D22)[A1,Dxe2x80x94P17,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 45);
Cyclo(K18xe2x80x94D22)[A1,Nle8,K18,D22,L27]hPTH(1-34)NH2 (SEQ ID NO: 46);
Cyclo(K18xe2x80x94D22)[A1,Nle8,D18,K22,L27]hPTH(1-31)NH2 (SEQ ID NO: 47);
Cyclo(O18xe2x80x94D22)[A1,Nle8,O18,O22,L27]hPTH(1-31)NH2 (SEQ ID NO: 48);
Cyclo(D18xe2x80x94O22)[A1,Nle8,D18,O22,L27]hPTH(1-31)NH2 (SEQ ID NO: 49);
Cyclo(K18xe2x80x94E22)[A1,Nle8,K18,E22,L27]hPTH(1-31)NH2 (SEQ ID NO: 50);
Cyclo(O18xe2x80x94E22)[A1,Nle8,O18,E22,L27]hPTH(1-31)NH2 (SEQ ID NO: 51);
Cyclo(K18xe2x80x94D22)[A1,Nle8,K18,D22,L27]hPTH(1-30)NH2 (SEQ ID NO: 52);
Cyclo(K18xe2x80x94D22)[A1,Nle8,K18,D22,L27]hPTH(1-29)NH2 (SEQ ID NO: 53);
Cyclo(K18xe2x80x94D22)[A1,Nle8,K18,D22,L27]hPTH(1-28)NH2 (SEQ ID NO: 54);
Cyclo(K18xe2x80x94D22)[A1,Nle8,K18,D22,L27]hPTH(1-27)NH2 (SEQ ID NO: 55);
Cyclo(K18xe2x80x94D22)[Nle8,K18,D22,L27]hPTH(3-31)NH2 (SEQ ID NO: 63);
Cyclo(K18xe2x80x94D22)[Nle8,K18,D22,L27]hPTH(2-31)NH2 (SEQ ID NO: 64);
Cyclo(K10xe2x80x94D14)[A1,Nle8,18,K10,D14,L27]hPTH(1-31)NH2 (SEQ ID NO: 66);
Cyclo(K14xe2x80x94D18)[A1,Nle8,K14,D18,L27]hPTH(1-31)NH2 (SEQ ID NO: 67);
Cyclo(K17xe2x80x94D21)[A1,Nle8,18,K17,D21,L27]hPTH(1-31)NH2 (SEQ ID NO: 68);
Cyclo(K21xe2x80x94D25)[A1,Nle8,18,K21,D25,L27]hPTH(1-31)NH2 (SEQ ID NO: 69);
Cyclo(K25xe2x80x94D29)[A1,Nle8,18,K25,D29,L27]hPTH(1-31)NH2 (SEQ ID NO: 70);
Cyclo(K18xe2x80x94D22)[K18,D22]hPTH(1-34)NH2 (SEQ ID NO: 71);
Cyclo(K18xe2x80x94D22)[K18,26,30,D22,L23,28,31,E25,29]hPTH(1-34)NH2 (SEQ ID NO: 72);
Bicyclo(K13xe2x80x94D17,K18xe2x80x94D22)[A1,Nle8,D17,22,K18,L27]hPTH(1-31)NH2 (SEQ ID NO: 73);
Bicyclo(K18xe2x80x94D22,K26xe2x80x94D30)[A1,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 74);
Tricyclo(K13xe2x80x94D17,K18xe2x80x94D22,K26xe2x80x94D30)[A1,Nle8,K18,D17,22,L27]hPTH(1-31)NH2 (SEQ ID NO: 80);
or a pharmaceutically acceptable salt or prodrug thereof.
Still more preferred cyclic peptide agonist compounds of this invention include:
Cyclo(K18xe2x80x94D22)[A1,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 3);
Cyclo(K18xe2x80x94D22)[A1,Nle8,K18,D22,L27]hPTH(1-34)NH2 (SEQ ID NO: 46);
Cyclo(K18xe2x80x94D22)[A1,3,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 6);
Cyclo(K18xe2x80x94D22)[A1,6,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 9);
Cyclo(K18xe2x80x94D22)[A1,10,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 12);
Cyclo(K18xe2x80x94D22)[A1,11,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 13);
Cyclo(K18xe2x80x94D22)[A1,12,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 14);
Cyclo(K18xe2x80x94D22)[A1,13,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 15);
Cyclo(K18xe2x80x94D22)[A1,14,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 16);
Cyclo(K18xe2x80x94D22)[A1,15,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 17);
Cyclo(K18xe2x80x94D22)[A1,16,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 18);
Cyclo(K18xe2x80x94D22)[A1,17,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 19);
Cyclo(K18xe2x80x94D22)[G1,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 20);
Cyclo(K18xe2x80x94D22)[A1,G2,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 21);
Cyclo(K18xe2x80x94D22)[A1,G3,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 22);
Cyclo(K18xe2x80x94D22)[A1,G10,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 29);
Cyclo(K18xe2x80x94D22)[A1,G13,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 31);
Cyclo(K18xe2x80x94D22)[A1,G16,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 34);
Cyclo(K18xe2x80x94D22)[A1,G17,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 35);
Cyclo(K18xe2x80x94D22)[Dxe2x80x94P1,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 36);
Cyclo(D18xe2x80x94K22)[A1,Nle8,D18,K22,L27]hPTH(1-31)NH2 (SEQ ID NO: 47);
Cyclo(O18xe2x80x94D22)[A1,Nle8,O18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 48);
Cyclo(K18xe2x80x94D22)[A1,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 49);
Cyclo(D18xe2x80x94O22)[A1,Nle8,D18,O22,L27]hPTH(1-31)NH2 (SEQ ID NO: 50);
Cyclo(K18xe2x80x94E22)[A1,Nle8,K18,E22,L27]hPTH(1-31)NH2 (SEQ ID NO: 51);
Cyclo(O18xe2x80x94E22)[A1,Nle8,O18,E22,L27]hPTH(1-30)NH2 (SEQ ID NO: 52);
Cyclo(K18xe2x80x94D22)[A1,Nle8,K18,D22,L27]hPTH(1-29)NH2 (SEQ ID NO: 53);
Cyclo(K18xe2x80x94D22)[A1,Nle8,K18,D22,L27]hPTH(1-28)NH2 (SEQ ID NO: 54);
Cyclo(K10xe2x80x94D14)[A1,Nle8,18,K10,D14,L27]hPTH(1-31)NH2 (SEQ ID NO: 66);
Cyclo(K14xe2x80x94D18)[A1,Nle8,K14,D18,L27]hPTH(1-31)NH2 (SEQ ID NO: 67);
Cyclo(K17xe2x80x94D21)[A1,Nle8,18,K17,D21,L27]hPTH(1-31)NH2 (SEQ ID NO: 68);
Cyclo(K21xe2x80x94D25)[A1,Nle8,18,K21,D25,L27]hPTH(1-31)NH2 (SEQ ID NO: 69);
Cyclo(K25xe2x80x94D29)[A1,Nle8,18,K25,D29,L27]hPTH(1-31)NH2 (SEQ ID NO: 70);
Cyclo(K18xe2x80x94D22)[K18,D22]hPTHrP(1-34)NH2 (SEQ ID NO: 71);
Cyclo(K18xe2x80x94D22)[K18,26,30,D22,L23,28,31,E25,29]hPTHrP(1-34)NH2 (SEQ ID NO: 72);
Bicyclo(K13xe2x80x94D17,K18xe2x80x94D22)[A1,Nle8,D17,22,K18,L27]hPTH(1-31)NH2 (SEQ ID NO: 73);
Bicyclo(K18xe2x80x94D22,K26xe2x80x94D30)[A1,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 74);
Tricyclo(K13xe2x80x94D17,K18xe2x80x94D22,K26xe2x80x94D30)[A1,Nle8,K18,D17,22,L27]hPTH(1-31)NH2 (SEQ ID NO: 80);
or a pharmaceutically acceptable salt or prodrug thereof.
Still yet more preferred cyclic peptide agonist compounds include:
Cyclo(K18xe2x80x94D22)[A1,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 3);
Cyclo(K18xe2x80x94D22)[A1,Nle8,K18,D22,L27]hPTH(1-34)NH2 (SEQ ID NO: 46);
Cyclo(K18xe2x80x94D22)[A1,10,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 12);
Cyclo(K18xe2x80x94D22)[A1,12,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 14);
Cyclo(K18xe2x80x94D22)[A1,13,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 15);
Cyclo(K18xe2x80x94D22)[A1,14,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 16);
Cyclo(K18xe2x80x94D22)[A1,16,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 18);
Cyclo(K18xe2x80x94D22)[A1,17,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 19);
Cyclo(K18xe2x80x94D22)[A1,G3,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 22);
Cyclo(K18xe2x80x94D22)[A1,G13,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 31);
Cyclo(K18xe2x80x94D22)[A1,G16,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 34);
Cyclo(K18xe2x80x94D22)[A1,G17,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 35);
Cyclo(K18xe2x80x94D22)[Dxe2x80x94P1,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 36);
Cyclo(D18xe2x80x94K22)[A1,Nle8,D18,K22,L27]hPTH(1-31)NH2 (SEQ ID NO: 47);
Cyclo(K18xe2x80x94E22)[A1,Nle8,K18,E22,L27]hPTH(1-31)NH2 (SEQ ID NO: 50);
Cyclo(O18xe2x80x94E22)[A1,Nle8,O18,E22,L27]hPTH(1-31)NH2 (SEQ ID NO: 51);
Cyclo(K18xe2x80x94D22)[A1,Nle8,K18,D22,L27]hPTH(1-30)NH2 (SEQ ID NO: 52);
Cyclo(K14xe2x80x94D18)[A1,Nle8,K14,D18,L27]hPTH(1-31)NH2 (SEQ ID NO: 67);
Cyclo(K18xe2x80x94D22)[K18,D22]hPTH(1-28)NH2 (SEQ ID NO: 71);
Bicyclo(K13xe2x80x94D17,K18xe2x80x94D22)[A1,Nle8,D17,22,K18,L27]hPTH(1-31)NH2 (SEQ ID NO: 73);
Bicyclo(K18xe2x80x94D22,K26xe2x80x94D30)[A1,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 74);
Tricyclo(K13xe2x80x94D17,K18xe2x80x94D22,K26xe2x80x94D30)[A1,Nle8,K18,D17,22,L27]hPTH(1-31)NH2 (SEQ ID NO: 80);
or a pharmaceutically acceptable salt or prodrug thereof.
Another still yet more preferred cyclic peptide agonist compound is
Bicyclo(K13xe2x80x94D17,K26xe2x80x94D30)[A1,Nle8,18,D17,L27]hPTH(1-31)NH2 (SEQ ID NO: 79)
or a pharmaceutically acceptable salt or prodrug thereof.
Certain cyclic peptide compounds of this invention inhibit the action of PTH. Such cyclic peptide antagonist compounds are useful in the treatment of disorders characterized by an excess of PTH such as hyperparathyrodism and hyperparathyrodism-related hypercalcemia crisis, hypercalcemia of malignancy, renal failure and hypertension.
A preferred cyclic peptide antagonist compound of formula 10 has formula 6 above wherein X is
(a) R1axe2x80x94A4xe2x80x94A5xe2x80x94A6xe2x80x94A7xe2x80x94A8xe2x80x94A9xe2x80x94,
(b) R1axe2x80x94A5xe2x80x94A6xe2x80x94A7xe2x80x94A8xe2x80x94A9xe2x80x94,
(c) R1axe2x80x94A6xe2x80x94A7xe2x80x94A8xe2x80x94A9xe2x80x94,
(d) R1axe2x80x94A7xe2x80x94A8xe2x80x94A9xe2x80x94,
(e) R1axe2x80x94A8xe2x80x94A9xe2x80x94,
(f) R1bxe2x80x94A9xe2x80x94, and
(g) R1bxe2x80x94;
A more preferred cyclic peptide antagonist compound of formula 11 has formula 10 above wherein A8 is Nle and A27 is Leu.
Another more preferred cyclic peptide antagonist compound of formula 12 has formula 11 above wherein wherein the side chains of A18 and A22 are linked through an amide bond to form a bridge.
Another more preferred cyclic peptide antagonist compound of formula 13 has formula 12 above wherein A10 is Asn or Asp; A13 is Lys; A14 is His or Ser; A17 is Asp or Ser; A18 is Asp, Lys or Orn; A21 is Arg or Val; A22 is Asp, Glu, Lys or Orn; A25 is Arg or His; A26 is His or Lys; A29 is Ala or Gln; and A30 is Asp or Glu.
More preferred cyclic peptide antagonist compounds include:
cyclo(K18xe2x80x94D22)[K18,D22,L27]hPTH(10-31)NH2 (SEQ ID NO: 56);
cyclo(K18xe2x80x94D22)[K18,D22,L27]hPTH(9-31)NH2 (SEQ ID NO: 57);
cyclo(K18xe2x80x94D22)[Nle8,K18,D22,L27]hPTH(8-31)NH2 (SEQ ID NO: 58);
cyclo(K18xe2x80x94D22)[Nle8,K18,D22,L27]hPTH(7-31)NH2 (SEQ ID NO: 59);
cyclo(K18xe2x80x94D22)[Nle8,K18,D22,L27]hPTH(6-31)NH2 (SEQ ID NO: 60);
cyclo(K18xe2x80x94D22)[Nle8,K18,D22,L27]hPTH(5-31)NH2 (SEQ ID NO: 61);
cyclo(K18xe2x80x94D22)[Nle8,K18,D22,L27]hPTH(4-31)NH2 (SEQ ID NO: 62);
cyclo(K18xe2x80x94D22)[Nle8,K18,D22,L27]hPTH(7-34)NH2 (SEQ ID NO: 65); and
cyclo(K18xe2x80x94D22)[K18,D22,]hPTH(7-34)NH2 (SEQ ID NO: 77);
or a pharmaceutically acceptable salt or prodrug thereof.
Certain acyclic peptide compounds of this invention also possess agonist activity on the parathyroid hormone receptor and accordingly are useful in the treatment of physiological conditions associated with bone cell calcium regulation including hypocalcemia; osteoporosis; osteopenia; and disorders associated with osteoporosis and osteopenia such as hyperparathyroidism, hypoparathyroidism, and Cushings syndrome; glucocorticoid- and immunosuppressant-induced osteopaenia; and bone fracture and bone refracture repair.
A preferred acyclic peptide agonist compound of formula 14 is the peptide compound of formula II wherein R1a is H and Y is NH2.
A more preferred acyclic peptide agonist compound of formula 15 has formula 14 above wherein X is
(a) R1axe2x80x94A1xe2x80x94A2xe2x80x94A3xe2x80x94A4xe2x80x94A5xe2x80x94A6xe2x80x94A7xe2x80x94A8xe2x80x94A9xe2x80x94,
(b) R1axe2x80x94A2xe2x80x94A3xe2x80x94A4xe2x80x94A5xe2x80x94A6xe2x80x94A7xe2x80x94A8xe2x80x94A9xe2x80x94 or
(c) R1axe2x80x94A3xe2x80x94A4xe2x80x94A5xe2x80x94A6xe2x80x94A7xe2x80x94A8xe2x80x94A9xe2x80x94,
Another more preferred acyclic peptide agonist compound of formula 16 has formula 15 above wherein A1 is Ser, Ala, Gly or D-Pro; A2 is Ala, Val or Gly; A3 is Ala, Ser, Gly or D-Pro; A4 is Glu, Ala or Gly; A5 is Ile, His, Ala or Gly; A6 is Ala, Gln, Gly or D-Pro; A7 is Ala, Leu, Gly; A8 is Leu, Nle, Gly or D-Pro; A9 is His, Ala, Gly or D-Pro; A10 is Ala, Asn, Gly, Asp or D-Pro; A11 is Ala, Gly, Leu or Lys; A12 is Ala or Gly; A13 is Ala, Gly or Lys; A14 is Ala, Gly, His, Ser or D-Pro; A15 is Ala, Gly, Ile or D-Pro; A16 is Asn, Ala, Gly, D-Pro or Gln; A17 is Ala, Asp, Gly, Ser or D-Pro; A18 is Lys; A19 is Arg or Glu; A20 is Arg; A21 is Arg or Val; A22 is Asp, Lys, Orn or Glu; A23 is Leu, Phe or Trp; A24 is Leu; A25 is Arg or His; A26 is Lys or His; A27 is Leu or Lys; A28 is Ile or Leu or an equivalent amino acid thereof; A29 is Ala or Gln; A30 is Asp or Glu; A3, is Ile, Leu or Val; A32 is His; A33 is Asn or Thr; and A34 is Ala or Phe.
Another more preferred acyclic peptide agonist compound of formula 17 has formula 16 above wherein A1 is Ala, Gly or D-Pro; A8 is Nle, A22 is Asp and A27 is Leu.
Another more preferred acyclic peptide agonist compound of formula 18 has formula 17 above wherein X is R1axe2x80x94A1xe2x80x94A2xe2x80x94A3xe2x80x94A4xe2x80x94A5xe2x80x94A6xe2x80x94A7xe2x80x94A8xe2x80x94A9xe2x80x94,
A still more preferred acyclic peptide agonist compound is [A1,Nle8,K18,D22,L27]hPTH(1-31)NH2 (SEQ ID NO: 4) or a pharmaceutically acceptable salt or prodrug thereof.
It is to be understood that th;s invention covers all appropriate combinations of the preferred aspects of the invention referred to herein.
The peptide compounds of the present invention may be synthesized by any techniques that are known to those skilled in the art of peptide synthesis. For solid phase peptide synthesis, a summary of the many techniques may be found in J. M. Stewart and J. D. Young, Solid Phase Peptide Synthesis, W. H. Freeman Co. (San Francisco), 1963 and J. Meienhofer, Hormonal Proteins and Peptides, vol. 2, p. 46, Academic Press (New York), 1973. For classical solution synthesis see G. Schroder and K. Lupke, The Peptides, vol. 1, Academic Press (New York), 1965.
In general, these methods comprise the sequential addition of one or more amino acids or suitably protected amino acids to a growing peptide chain. Normally, either the amino or carboxyl group of the first amino acid is protected by a suitable protecting group. The protected or derivatized amino acid can then either be attached to an inert solid support or utilized in solution by adding the next amino acid in the sequence having the complimentary (amino or carboxyl) group suitably protected, under conditions suitable for forming the amide linkage. The protecting group is then removed from this newly added amino acid residue and the next amino acid (suitably protected) is then added, and so forth. After all the desired amino acids have been linked in the proper sequence, any remaining protecting groups (and any solid support) are removed sequentially or concurrently, to afford the final peptide compound. By simple modification of this general procedure, it is possible to add more than one amino acid at a time to a growing chain, for example, by coupling (under conditions which do not racemize chiral centers) a protected tripeptide with a properly protected dipeptide to form, after deprotection, a pentapeptide and so forth.
A preferred method of preparing the peptide compounds of the present invention involves solid phase peptide synthesis.
In this particularly preferred method the alpha-amino function is protected by an acid or base sensitive group. Such protecting groups should have the properties of being stable to the conditions of peptide linkage formation, while being readily removable without destruction of the growing peptide chain or racemization of any of the chiral centers contained therein. Suitable protecting groups are 9-fluorenylmethyloxycarbonyl (Fmoc), t-butyloxycarbonyl (Boc), benzyloxycarbonyl (Cbz), biphenylisopropyloxycarbonyl, t-amyloxycarbonyl, isobornyloxycarbonyl, (xcex1,xcex1)dimethyl-3,5-dimethoxybenzyloxycarbonyl, o-nitrophenylsulfenyl, 2-cyano-t-butyloxycarbonyl, and the like. The 9-fluorenylmethyloxycarbonyl (Fmoc) protecting group is preferred.
Particutlarly preferred side chain protecting groups are, for side chain amino groups as in lysine and arginine: 2,2,5,7,8-pentamethylchroman-6-sultonyl (pmc), nitro, p-toluenesulfonyl, 4-methoxybenzenesulfonyl, Cbz, Boc, Alloc (allyloxycarbonyl) and adamantyloxycarbonyl; for tyrosine: benzyl, o-bromobenzyloxycarbonyl, 2,6-dichlorobenzyl, isopropyl, t-butyl (t-Bu), cyclohexyl, cyclopenyl and acetyl (Ac); for serine: t-butyl, benzyl and tetrahydropyranyl; for histidine: trityl, benzyl, Cbz, p-toluenesulfonyl and 2,4-dinitrophenyl; for tryptophan: formyl and Boc; for asparagine and glutamine: Trt (trityl); for aspartic acid and glutamic acid: O-t-Bu and OAllyl.
The cyclic peptide compounds of this invention are preferably prepared using a fragment-based approach in which a fragment of the desired complete peptide compound containing the amide, ester, disulfide or lanthionine bridge (the cyclic peptide fragment) is separately prepared and purified prior to coupling to a resin-bound amino acid or peptide portion of the complete peptide compound. The cyclic peptide fragment is prepared using classical solution phase synthetic techniques or solid phase peptide synthesis methodology as described herein. The synthesis of the complete peptide compound is then accomplished by sequential addition of the remaining amino acid residues to the resin-bound cyclic peptide; by addition of additional cyclic or acyclic peptide fragments to the resin bound cyclic peptide; or by any combination of the above. The preparation of cyclic peptide hPTH analogs using a fragment-based approach is described in U.S. Ser. No. 60/081897, filed Apr. 15, 1998, incorporated herein by reference.
Peptides of this invention wherein R1c is 
are prepared using the method described by R. Waelchli et al., Tetrahedron Lett., 6(10), 1151-1156 (1996), incorporated herein by reference.